Pure and Zr-doped NiO nanoparticles were successfully synthesized through green synthesis using extract of Annona squamosa seeds. The synthesized nanoparticles were characterized by X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, UV-vis spectroscopy, field emission scanning electron microscopy (FESEM), EDAX (energy dispersive X-ray analysis) and elemental mapping analysis. The addition of zirconium dopant has been integrated into the single-phase NiO lattice structure as indicated by characterization findings. Furthermore, the antibacterial, antifungal and antioxidant properties of Zr-doped and undoped NiO NPs were thoroughly examined. As a result of the strong Zr-doped NiO NPs synergistic effect, the results showed that the Zr-doped NiO NPs have improved the biomedical properties when compared to the undoped NiO NPs. The optimized size and shape of the nanoparticles contributed significantly to their improved effectiveness against the microorganisms and oxidative stress.